Evaluation of the genotoxicity of neurotoxic insecticides using the micronucleus test in Tradescantia pallida.

Neonicotinoids and phenylpyrazoles are classes of neurotoxic insecticides which are able to bind at different ligand sites of neural receptors, leading to the deregulation of insect neural activity and hence resulting in death. The misuse or indiscriminate use of these chemicals is directly associated with several toxicological effects in biota and at different trophic levels. Based on this premise, the aim of the present study was to evaluate and compare the genotoxic capacity of different concentrations of thiamethoxam (TMX), acetamiprid (ACP), imidacloprid (IMI) and fipronil (FP) through the Micronucleus Test in Tradescantia pallida (Trad-MCN). After acclimatization (24 h), T. pallida stems were treated with stablished concentrations of TMX, ACP, IMI and FP for 8 h. Then, the stems of the model organism were submitted to a recovery phase (24 h). The young inflorescences were harvested and fixed in Carnoy solution and, after 24 h, were conserved in ethanol 70% until the analyzes. The obtained anthers were macerated on slides for microscopy, stained with acetic carmine dye and covered with coverslips before analysis by light microscopy. Considering the insecticides, the micronuclei (MN) frequency in plants treated at concentrations of 0.2 and 0.4 g L-1 for TMX, 0.2; 0.4 and 0.8 g L-1 for ACP, 0.1; 0.2; 0.4; 0.8 and 1.6 g L-1 for IMI and 0.2; 0.4; 0.8 and 1.6 g L-1 for FP differed statistically (p < 0.05, Tukey) from the MN frequency of the negative control. All chemicals evaluated revealed genotoxic activity in T. pallida at the highest concentrations.

Floral visitors of canola (Brassica napus L. ) hybrids in Cerrado Mineiro region, Brazil / Visitantes florais em híbridos de canola (Brassica napus L. ) na região do Cerrado Mineiro, Brasil

The objective of this study was to verify floral visitors of canola in the Triângulo Mineiro region, Minas Gerais, Brazil. A completely randomized plot design with two hybrids and 10 replicates was used, totaling 20 plots. The survey of floral visitors was made during 10 days. Faunal analyses of the collected species were performed based on frequency, constancy, dominance, and diversity. A total of 2,676 individuals belonging to 13 species of bees of two families (Apidae and Halictidae) were collected. The diversity index found among hybrids was similar (1.28 in hybrid 61 and 1.50 in hybrid 433). The species Apis mellifera Linnaeus, 1758 (Hymenoptera: Apidae), Paratrigona lineata (Lepeletier, 1836) (Hymenoptera: Apidae), and Trigona spinipes (Fabricius, 1793) (Hymenoptera: Apidae) were the most abundant and constant in the crops during all the evaluated days for both hybrids. So, it is important that efforts be made to conserve natural areas close to crops in order to provide resources for the permanence of floral visitors in the area.(AU)

O objetivo deste estudo foi verificar os visitantes florais de canola na região do Triângulo Mineiro, Minas Gerais, Brasil. ­Utilizou-se um delineamento experimental de blocos inteiramente randomizados com dois híbridos e 10 amostras replicadas, totalizando 20 amostras. A pesquisa de visitantes florais foi feita durante 10 dias. As análises faunísticas das espécies coletadas foram realizadas com base na frequência, constância, domínio, riqueza e diversidade. Um total de 2.676 indivíduos pertencentes a 13 espécies de abelhas de duas famílias (Apidae e Halictidae) foram coletados. A riqueza das espécies encontradas entre os híbridos foi semelhante (10 espécies no híbrido 61 e 12 espécies no híbrido 433), bem como o índice de diversidade (1,28 no híbrido 61 e 1,50 no híbrido 433). A espécie Apis mellifera Linnaeus, 1758 (Hymenoptera: Apidae), Paratrigona lineata (Lepeletier, 1836) (Hymenoptera: Apidae) e Trigona spinipes (Fabricius, 1793) (Hymenoptera: Apidae) foram as mais abundantes e constantes nas culturas durante todos os dias avaliados para ambos os híbridos. Com isso, é importante que sejam feitos esforços para conservar as áreas naturais próximas às culturas, a fim de fornecer recursos para a permanência de visitantes florais na área.(AU)

Ecotoxicological effects of the insecticide fipronil in Brazilian native stingless bees Melipona scutellaris (Apidae: Meliponini).

Melipona scutellaris Latreille, 1811 (Hymenoptera, Apidae) is a pollinator of various native and cultivated plants. Because of the expansion of agriculture and the need to ensure pest control, the use of insecticides such as fipronil (FP) has increased. This study aimed to evaluate the effects of sublethal doses of FP insecticide on M. scutellaris at different time intervals (6, 12, and 24 h) after exposure, via individually analyzed behavioral biomarkers (locomotor activity, behavioral change) as well as the effect of FP on different brain structures of bees (mushroom bodies, antennal cells, and optic cells), using sub-individual cell biomarkers (heterochromatin dispersion, total nuclear and heterochromatic volume). Forager bees were collected when they were returning to the nest and were exposed to three different concentrations of FP (0.40, 0.040, and 0.0040 ng a.i/bee) by topical application. The results revealed a reduction in the mean velocity, lethargy, motor difficulty, paralysis, and hyperexcitation in all groups of bees treated with FP. A modification of the heterochromatic dispersion pattern and changes in the total volume of the nucleus and heterochromatin were also observed in the mushroom bodies (6, 12, and 24 h of exposure) and antennal lobes (6 and 12 h) of bees exposed to 0.0040 ng a.i/bee (LD50/100). FP is toxic to M. scutellaris and impairs the essential functions required for the foraging activity.

Ocimum basilicum essential oil combined with deltamethrin to improve the management of Spodoptera frugiperda / Óleo essencial de Ocimum basilicum associado à deltametrina no manejo de Spodoptera frugiperda

ABSTRACT For an important and expensive crop such as corn, the resistance of Spodoptera frugiperda J.E. Smith to various pesticides has led to research throughout the world for a potential insecticide from a natural source. For the management of pest resistance, natural compounds associated with synthetic insecticides can be a promising tool because they can reduce the application of the synthetics molecules while maintaining their effectiveness and promoting the control of the pests. Linalool is a potential insecticide that is easily obtained because it is found in high concentrations in the essential oil of Ocimum basilicum L. Therefore, the present study aimed to evaluate the toxicity of this essential oil and its combination with deltamethrin to control S. frugiperda. Through dose response assays, the acute toxicities (LD50) of the essential oil and deltamethrin were estimated. Additionally, the combination of these materials was also assessed, attaining a reduction of 80% of the LD50 of deltamethrin while obtaining the same result as when the pyrethroid was administered alone. From these results, it is expected that the combination of natural compounds and synthetic insecticides will be a promising practice, helping to manage resistance while reducing the environmental impact of toxic compounds.

RESUMO Para uma cultura importante e expressiva como o milho, a resistência de Spodoptera frugiperda J.E. Smith a vários inseticidas atraiu a atenção no mundo para pesquisar o potencial inseticida de compostos naturais. Para o manejo da resistência, os compostos naturais associados a inseticidas sintéticos podem ser uma ferramenta promissória por reduzirem a aplicação das moléculas sintéticas para que não percam sua eficácia, além de promover o controle das pragas. O linalol é um terpenoide considerado como inseticida potencial, pode ser facilmente obtido naturalmente uma vez que é encontrado em alta concentração no óleo essencial de Ocimum basilicum. Portanto, o presente estudo teve como objetivo avaliar a toxicidade deste óleo essencial e sua combinação com deltametrina no controle de Spodoptera frugiperda. Através do ensaio de dose-resposta, foi estimada a toxicidade aguda (DL50) do óleo essencial e da deltametrina. Adicionalmente, foi também avaliada a combinação entre ambos, alcançando uma redução de 80% da DL50 de deltametrina para se obter o mesmo resultado quando o piretroide foi administrado sozinho. A partir dos nossos resultados, espera-se que uma combinação de uso de compostos naturais e inseticidas sintéticos possa ser uma prática promissora, auxiliando no manejo da resistência de pragas e principalmente reduzindo os impactos ambientais de compostos tóxicos.

Mutagenic, recombinogenic and carcinogenic potential of thiamethoxam insecticide and formulated product in somatic cells of Drosophila melanogaster.

Thiamethoxam (TMX) belongs to a class of neuro-active insecticides referred as neonicotinoids, while actara® (AC) is one of the most popular TMX-based products in Brazil. The aim of this study was to evaluate the mutagenic, recombinogenic and carcinogenic potential of TMX and AC insecticides. The mutagenic and recombinogenic effect of TMX and AC were evaluated in vivo by the Somatic Mutation and Recombination Test (SMART) while carcinogenic effects were evaluated through the Test for Detection of Epithelial Tumor Clones (wts test), both in somatic cells of Drosophila melanogaster. In the SMART, third instar larvae from standard (ST) and high bioactivation (HB) crosses were treated with different concentrations of TMX and AC (2.4; 4.8; 9.7 × 10-4 mM and 1.9 × 10-3 mM). The results revealed mutagenic effects at the highest concentrations tested in the HB cross. In the test for the detection of epithelial tumor, third instar larvae resulting from the cross between wts/TM3, Sb1 virgin females and mwh/mwh males were treated with the same concentrations of TMX and AC used in the SMART. No carcinogenic effect was observed at any of the concentrations tested. In this work, the inhibition of the mechanism of repair by homologous recombination was observed in flies exposed to 9.7 × 10-4 and 1.9 × 10-3 mM of AC. In conclusion, TMX and AC demonstrated to be a promutagen in the highest concentrations tested.

Exposure of larvae to thiamethoxam affects the survival and physiology of the honey bee at post-embryonic stages.

Under laboratory conditions, the effects of thiamethoxam were investigated in larvae, pupae and emerging honey bees after exposure at larval stages with different concentrations in the food (0.00001 ng/µL, 0.001 ng/µL and 1.44 ng/µL). Thiamethoxam reduced the survival of larvae and pupae and consequently decreased the percentage of emerging honey bees. Thiamethoxam induced important physiological disturbances. It increased acetylcholinesterase (AChE) activity at all developmental stages and increased glutathione-S-transferase (GST) and carboxylesterase para (CaEp) activities at the pupal stages. For midgut alkaline phosphatase (ALP), no activity was detected in pupae stages, and no effect was observed in larvae and emerging bees. We assume that the effects of thiamethoxam on the survival, emergence and physiology of honey bees may affect the development of the colony. These results showed that attention should be paid to the exposure to pesticides during the developmental stages of the honey bee. This study represents the first investigation of the effects of thiamethoxam on the development of A. mellifera following larval exposure.

Assessment of the mutagenic, recombinogenic and carcinogenic potential of fipronil insecticide in somatic cells of Drosophila melanogaster.

Fipronil (FP) is an insecticide that belongs to the phenylpyrazole chemical family and is used to control pests by blocking GABA receptor at the entrance channel of the chlorine neurons. The aim of this study was to evaluate the mutagenic, recombinogenic and carcinogenic potential of FP. The mutagenic and recombinogenic effects were evaluated using the somatic mutation and recombination test (SMART) on wing cells of Drosophila melanogaster. Third instar larvae from standard (ST) and high bioactivation (HB) crosses were treated with different concentrations of FP (0.3, 0.7, 1.5 or 3.0 × 10-5 mM). The results showed mutagenic effects at all concentrations tested in the HB cross; and all concentrations tested in the ST cross, except at concentration of 0.7 × 10-5 mM. The carcinogenic effect of FP was assayed through the test for detection of epithelial tumor (warts) in D. melanogaster. Third instar larvae from wts/TM3 virgin females mated to mwh/mwh males were treated with different concentrations of FP (0.3, 0.7, 1.5 or 3.0 × 10-5 mM). All these concentrations induced a statistically significant increase in tumor frequency. In conclusion, FP proved to be mutagenic, recombinogenic and carcinogenic in somatic cells of D. melanogaster.

Assessment of water quality and genotoxic impact by toxic metals in Geophagus brasiliensis.

This study used the pearl cichlid Geophagus brasiliensis as a bioindicator to survey the health of the aquatic environment on four sites (P1, P2, P3 and P4) of the Mumbuca stream located at Monte Carmelo/MG, Brazil. The selection of different sites was made with reference to the gradient of urban activity and via physicochemical and biological evaluation of water quality and genotoxicity. The water quality index was classified as 'good' for P1 and P4, regular in P2 and 'poor' for P3. The micronuclei (MN) frequency obtained from blood analysis was in agreement with the water quality, such that the higher values of MN were detected in sites evaluated as poor. Water degradation conditions worsen according to the flow of the stream over the sites P1, P2 and P3, but for site P4, located after the Monte Carmelo Sewage Treatment Plant, improvements in the micronuclei frequency are detected. Our results showed high levels of potentially toxic metals (chromium, lead, aluminum and nickel) in specific stream sites (P2 and P3). We suggest that the micronuclei induction in G. brasiliensis could be due to the presence of these compounds.

Effects of Nosema ceranae and thiametoxam in Apis mellifera: A comparative study in Africanized and Carniolan honey bees.

Multiple stressors, such as chemicals and pathogens, are likely to be detrimental for the health and lifespan of Apis mellifera, a bee species frequently exposed to both factors in the field and inside hives. The main objective of the present study was to evaluate comparatively the health of Carniolan and Africanized honey bees (AHB) co-exposed to thiamethoxam and Nosema ceranae (N. ceranae) spores. Newly-emerged worker honey bees were exposed solely with different sublethal doses of thiamethoxam (2% and 0.2% of LD50 for AHB), which could be consumed by bees under field conditions. Toxicity tests for the Carniolan bees were performed, and the LD50 of thiamethoxam for Carniolan honey bees was 7.86 ng bee(-1). Immunohistological analyses were also performed to detect cell death in the midgut of thiamethoxam and/or N. ceranae treated bees. Thiamethoxam exposure had no negative impact on Nosema development in experimental conditions, but it clearly inhibited cell death in the midgut of thiamethoxam and Nosema-exposed bees, as demonstrated by immunohistochemical data. Indeed, thiamethoxam exposure only had a minor synergistic toxic effect on midgut tissue when applied as a low dose simultaneously with N. ceranae to AHB and Carniolan honey bees, in comparison with the effect caused by both stressors separately. Our data provides insights into the effects of the neonicotenoid thiamethoxam on the AHB and Carniolan honey bee life span, as well as the effects of simultaneous application of thiamethoxam and N. ceranae spores to honey bees.

In vitro effects of thiamethoxam on larvae of Africanized honey bee Apis mellifera (Hymenoptera: Apidae).

Several investigations have revealed the toxic effects that neonicotinoids can have on Apis mellifera, while few studies have evaluated the impact of these insecticides can have on the larval stage of the honeybee. From the lethal concentration (LC50) of thiamethoxam for the larvae of the Africanized honeybee, we evaluated the sublethal effects of this insecticide on morphology of the brain. After determine the LC50 (14.34 ng/µL of diet) of thiamethoxam, larvae were exposed to a sublethal concentration of thiamethoxam equivalent to 1.43 ng/µL by acute and subchronic exposure. Morphological and immunocytochemistry analysis of the brains of the exposed bees, showed condensed cells and early cell death in the optic lobes. Additional dose-related effects were observed on larval development. Our results show that the sublethal concentrations of thiamethoxam tested are toxic to Africanized honeybees larvae and can modulate the development and consequently could affect the maintenance and survival of the colony. These results represent the first assessment of the effects of thiamethoxam in Africanized honeybee larvae and should contribute to studies on honey bee colony decline.

Toxicity of Imidacloprid to the Stingless Bee Scaptotrigona postica Latreille, 1807 (Hymenoptera: Apidae).

The stingless bee Scaptotrigona postica is an important pollinator of native and cultivated plants in Brazil. Among the factors affecting the survival of these insects is the indiscriminate use of insecticides, including the neonicotinoid imidacloprid. This work determined the toxicity of imidacloprid as the topical median lethal dose (LD50) and the oral median lethal concentration (LC50) as tools for assessing the effects of this insecticide. The 24 and 48 h LD50 values were 25.2 and 24.5 ng of active ingredient (a.i.)/bee, respectively. The 24 and 48 h LC50 values were 42.5 and 14.3 ng a.i./µL of diet, respectively. Ours results show the hazard of imidacloprid and the vulnerability of stingless bees to it, providing relevant toxicological data that can used in mitigation programs to ensure the conservation of this species.

Composição química e toxicidade de óleos essenciais para o pulgão-verde Schizaphis graminum (Rondani, 1852) / Chemical composition and toxicity of essential oils to the green-aphid Schizaphis graminum (Rondani, 1852)

Este trabalho teve como objetivo identificar e quantificar os constituintes dos óleos essenciais de Illicum verum, Ageratum conyzoides, Piper hispidinervum e Ocotea odorifera, bem como avaliar a toxicidade para o pulgão-verde Schizaphis graminum. A qualificação dos constituintes foi realizada por meio de um cromatógrafo gasoso + espectrômetro de massas, e a quantificação, por um cromatógrafo gasoso + detector de ionização de chama, ambos com uma coluna DB5. O método de hidrodestilação promoveu um rendimento (p/p) de 3,81% para I. verum, 0,46% para A. conyzoides, 2,85% para P. hispidinervum e 0,68% para O. odorífera. Já os componentes majoritários foram: precoceno (87,0%) e (E)-cariofileno (7,1%) para A. conyzoides; (E)-anetol (90,4%), limoneno (2,6%) e metil-chavicol (1,3%) para I. verum; metil-eugenol (81,2%) e safrol (10,6%) para Ocotea odorífera; e safrol (82,5%) e α-terpinoleno (13,4%) para P. hispidinervum. Pelos testes de toxicidade aguda (24 horas) com folhas de sorgo ou papel-filtro contaminados, verificou-se que o óleo de A. conyzoides foi o mais tóxico para o pulgão, com CL50 de 7,13 e 7,08 µL óleo/cm2 respectivamente, seguido por O. odorifera com CL50 de 11,80 e 103,00 µL óleo/cm2 respectivamente; I. verum de 51,80 µL óleo/cm2 em ambos os substratos; e o menos tóxico foi o óleo essencial de P. hispidinervum, com CL50 de 62,50 e 143,00 µL óleo/cm2, respectivamente. Dessa maneira, sugere-se que o uso dos óleos essenciais pode representar uma nova ferramenta em programas de manejo integrado de pragas.(AU)

The aim of this study was to assess the chemical composition of essential oils of Illicum verum, Ageratum conyzoides, Piper hispidinervum and Ocotea odorífera, as well as their toxicity to the green-aphid Schizaphis graminum. Compound identification was carried out with gas chromatography + mass spectrometry, and quantification with gas chromatography + flame ionization detector, both with DB5 column. The hydrodistillation process promoted oil yield of 3.81% for I. verum; 0.46% for A. conyzoides; 2.85 % for P. hispidinervum; and 0.68 % for O. odorífera. The major components from the oils of A. conyzoides was precocene (87.0%) and (E)-caryophyllene (7.1%); (E)-anethol (90.4%), limonene (2.6%) and methyl-chavicol (1.3%) for I. verum; methyleugenol (81.2%) and safrole (10.6%) for O. odorifera; and safrole (82.5%) and α-terpinolene (13.4%) for P. hispidinervum. The acute toxicity test (24 hours) performed by contact in sorghum leaves or contaminated filter-paper showed that the higher toxicity occur with essential oils of A. conyzoides, with LC50 of 7.01 and 7.01 µg oil/cm2, respectively; followed by O. odorifera, with LC50 of 17.10 and 66.70 µg oil/cm2, respectively; I. verum, with LC50of 66.40 and 65.40 µg oil/cm2, respectively; and low toxicity was observed for the P. hispidinervum essential oil, with LC50 of 64.00 and 143.00 oil/cm2, respectively. With the results obtained in this research, we can state that the use of these essential oils can be a new tool in integrated pest management.(AU)

Side-effects of thiamethoxam on the brain andmidgut of the africanized honeybee Apis mellifera (Hymenopptera: Apidae).

The development of agricultural activities coincides with the increased use of pesticides to control pests, which can also be harmful to nontarget insects such as bees. Thus, the goal of this work was assess the toxic effects of thiamethoxam on newly emerged worker bees of Apis mellifera (africanized honeybee-AHB). Initially, we determined that the lethal concentration 50 (LC50 ) of thiamethoxam was 4.28 ng a.i./µL of diet. To determine the lethal time 50 (LT50 ), a survival assay was conducted using diets containing sublethal doses of thiamethoxam equal to 1/10 and 1/100 of the LC50. The group of bees exposed to 1/10 of the LC50 had a 41.2% reduction of lifespan. When AHB samples were analyzed by morphological technique we found the presence of condensed cells in the mushroom bodies and optical lobes in exposed honeybees. Through Xylidine Ponceau technique, we found cells which stained more intensely in groups exposed to thiamethoxam. The digestive and regenerative cells of the midgut from exposed bees also showed morphological and histochemical alterations, like cytoplasm vacuolization, increased apocrine secretion and increased cell elimination. Thus, intoxication with a sublethal doses of thiamethoxam can cause impairment in the brain and midgut of AHB and contribute to the honeybee lifespan reduction.

Acute toxicity of fipronil to the stingless bee Scaptotrigona postica Latreille.

Fipronil is an insecticide widely used to control a great number of pests, thus the aim of this study was to determine the lethal dose and lethal concentration (LD(50) and LC(50)) of this insecticide to the stingless bees Scaptotrigona postica Latreille, 1807. The LD(50) and LC(50) values obtained after 24 h of exposition were of 0.54 ng a.i./bee and 0.24 ng a.i./µL diet, respectively. These values were considered highly toxic to stingless bees.

Oral toxicity of fipronil insecticide against the stingless bee Melipona scutellaris (Latreille, 1811).

For a better evaluation of the model using Apis mellifera in toxicology studies with insecticides, the oral acute toxicity of the insecticide fipronil against the stingless bee Melipona scutellaris was determined. The results showed that fipronil was highly toxic to M. scutellaris, with a calculated LC(50) (48 h) value of 0.011 ng a.i./µL of sucrose solution and an estimated oral LD(50) (48 h) of 0.6 ng a.i./bee. Our results showed that M. scutellaris bee is more sensitive to fipronil than the model specie A. mellifera.

Chemical composition and fumigant effect of essentialoil of Lippia sidoides Cham. and monoterpenes against Tenebrio molitor (L.) (coleoptera: tenebrionidae) / Composição química e efeito fumigante do óleo essencial de Lippia sidoides Cham. e monoterpenos sobre Tenebrio molitor (L.) (Coleoptera: Tenebrionidae)

The chemical composition of Lippia sidoides essential oils obtained by hydrodistillation was characterized and quantified by GC/MS and their insecticidal activity by fumigation test was assayed against Tenebrio molitor. Moreover, the toxicity of monoterpenes carvacrol, 1,8-cineol and thymol were also evaluated when applied alone or in binary (1:1) or tertiary (1:1:1) mixture. The essential oil of L. sidoides has as major constituents carvacrol (31.68 percent), ρ-cymene (19.58 percent), 1,8-cineole (9.26 percent) and ϒ-terpinene (9.21 percent), from a 21 compounds identified, being 92.53 percent of total. Both compounds have insecticidal activity against T. molitor, being the degree of toxicity of carvacrol > 1,8-cineole > L. sidoides essential oil > thymol, and its respectively LC50 at 24h were 5.53; 5.71; 8.04 and 14.71 µL/L air. When the different mixture of carvacrol, 1,8-cineole and thymol was assayed against T. molitor, the synergism among them was observed. For the mixture of carvacrol:1,8-cineole LC50 was 5.34 µL/L air; carvacrol:thymol 7.67 µL/L air; 1,8-cineole:thymol 7.51 µL/L air and carvacrol:1,8-cineole:thymol 6.34 µL/L air. Mainly, the monoterpene thymol had a synergic effect, which increased the toxicity of carvacrol and 1,8-cineole, both in binary mixture like carvacrol:thymol and 1,8-cineole:thymol.

A composição química do óleo essencial de Lippia sidoides obtido por hidrodestilação foi caracterizada e quantificada por GC/MS, bem como sua atividade inseticida por teste de fumigação foi avaliada sobre Tenebrio molitor. Além disso, a toxicidade dos monoterpenos carvacrol, 1,8-cineol e timol, também foi avaliada quando esses compostos foram aplicados isoladamente, ou em misturas binárias (1:1), ou terciárias (1:1:1). O óleo essencial de L. sidoides tem como principais constituintes o carvacrol (31,68 por cento), ρ-cimeno (19,58 por cento), 1,8-cineol (9,26 por cento) e ϒ-terpineno (9,21 por cento), em 21 compostos identificados, sendo 92,53 por cento do total. Ambos os compostos possuem atividade inseticida contra T. molitor, seguindo a ordem de toxicidade: carvacrol > 1,8-cineol > óleo essencial de L. sidoides > timol; sendo suas respectivas CL50 à 24 h de 5,53; 5,71; 8,04 e 14,71 µL/L ar. Quando as diferentes misturas de carvacrol, 1,8-cineol e timol foram avaliadas contra T. molitor, verificou-se o efeito sinérgico. Para a mistura de carvacrol:1,8-cineol a CL50 foi de 5,34 µL/L ar; carvacrol:timol de 7,67 µL/L ar; 1,8-cineol:timol de 7,51 µL/L ar e carvacrol:1,8-cineol:timol de 6,34 µL/L ar. Principalmente o monoterpeno timol teve efeito sinérgico, aumentando a toxicidade de carvacrol e 1,8-cineol quando em misturas binárias, tais como carvacrol:timol e 1,8-cineol:timol.

Inversão da sacarose utilizando ácido cítrico e suco de limão para preparo de dieta energética de Apis mellifera Linnaeus, 1758 / Inversion of the sucrose using citric acid and lemon juice for preparing energetic diet of Apis mellifera Linnaeus, 1758

Em época de escassez de néctar, pode-se fornecer ao enxame uma suplementação alimentar utilizando "açúcar invertido" que é obtido pela hidrólise da sacarose em meio ácido por aquecimento, formando uma mistura de glicose e frutose. O ácido normalmente utilizado em tal reação é o ácido cítrico, no entanto, diante da dificuldade de pequenos produtores apícolas em obterem tal produto comercialmente, uma alternativa seria substituí-lo pela utilização de suco de limões. Sendo assim, o objetivo deste trabalho foi quantificar a inversão da sacarose em dietas para operárias de Apis mellifera, empregando-se o ácido cítrico e sucos de limões. Prepararam-se 21 soluções aquosas de açúcar cristal na proporção de 100 g/100 mL, colocando-se em cada uma 1,0; 2,0; 3,0; 5,0; 10,0; 15,0 ou 20,0 mL de suco dos limões Galego [Citrus aurantifolia (C.) Swingle], Tahiti [Citrus latifolia Tanaka] ou Cravo [Citrus limonia (L.) Osbeck]. Além dessas, outras seis soluções aquosas de açúcar na mesma proporção foram preparadas adicionando-se 0,1; 0,16; 0,3; 0,5 e 0,7 g de ácido cítrico e uma para testemunha. Quantificou-se a inversão pelo método de Somogyi-Nelson. Para o limão Cravo foi encontrada a menor porcentagem de inversão. Em relação a adição de ácido cítrico, constatou-se que ao colocar 0,1 g, a inversão foi estimada em 12,2 por cento enquanto que, com a adição de 0,16 g, a inversão seria de 18,8 por cento. A quantidade máxima de ácido cítrico a ser adicionada foi estimada em 0,18 g adotando como critério o pH médio de 3,3 dos méis de abelhas africanizadas. Para os sucos dos limões Galego, Tahiti e Cravo, quando utilizados como substitutos do ácido cítrico, estimou-se a quantidade máxima a ser adicionada sendo igual a 2,1; 3,6 e 5,3 mL, respectivamente, para cada 100 g de açúcar em 100 mL de água, para o pH fixado em 3,3. Considerando-se a inversão e o pH, o suco do limão Tahiti forneceu melhores resultados.

In times of nectar shortage, food supplementation using "inverted sugar" obtained by sucrose hydrolysis in an acid environment by heating can be supplied to the hive, forming a glucose and fructose mixture. The acid usually used in such a reaction is the citric acid, however, due to the difficulty that small producers have to obtain the product commercially, an alternative would be to replace it with lemon juice. The objective of this work was to quantify the inversion of sucrose present in Apis mellifera worker diets by employing commercial citric acid and lemon juice. A total of 21 aqueous solutions of granulated sugar at the proportion of 100 g/100 mL were prepared, placing in each one 1.0; 2.0; 3.0; 5.0; 10.0; 15.0 or 20.0 mL of the juices of the Galego [Citrus aurantifolia (C.) Swingle], Tahiti [Citrus latifolia Tanaka] or Cravo [Citrus limonia (L.) Osbeck] lemon varieties. In addition to those, six other aqueous sucrose solutions were prepared by adding 0.0 (control); 0.16; 0.3; 0.5 and 0.7 g citric acid. Sucrose inversion was quantified by using the Somogyi-Nelson method. For the Cravo lemon, the lowest inversion rate was found. Regarding the addition of citric acid, it was estimated that in placing 0.1 g, the inversion was of 12.2 percent while with 0.16 g, an inversion of 18.8 percent sucrose present in the solution. The maximum amount of citric acid to be added was estimated to be 0.18 g, adopting as a criterion the mean pH of 3.3 in Africanized bee honey. If Galego, Tahiti and Cravo lemon juices are used as replacements for the citric acid, the maximum added amount should be 2.1, 3.6 and 5.3 mL, respectively, for each 100 g sugar/100 mL water, at fixed pH of 3.3. Considering the inversion rate and pH level recommended for the use of citric acid, Tahiti lemon juice provides better results.

Calli induction in leaf explants of coffee elite genotypes / Indução de calos em explantes foliares de clones elite de café

Three experiments were carried out with the objective of achieving high effectiveness in calli induction from high heterozygosis leaf explants of Coffea arabica through indirect somatic embryogenesis. A randomized-block design in a 2x5 factorial arrangement made up of two media [BOXTEL & BERTHOULY (1996) and TEIXEIRA et al. (2004)] and five C. arabica genotypes were used in the first experiment. In the second experiment the embryogenic calli production potential was evaluated in ten genotypes. Each of them was considered as a treatment. In the third experiment the variations in both 2.4-D (2.5 e 20µM) and 2-iP (2.5 e 20µM) concentrations in TEIXEIRA et al. (2004) medium and secondary media were evaluated. Crops were kept in a growth room under darkness, at 25±2oC. The medium described by TEIXEIRA et al (2004) was found to be superior when compared to that described by BOXTEL & BERTHOULY (1996) in the 2.2 and 7.2 genotypes. An opposite behavior was noticed in 4.2 genotype, that is, BOXTEL & BERTHOULY (1996) had medium superiority. Both 3.0 and 5.0 genotypes had the same behavior in both media studied, which shows that the somatic embryo production depends on the genotype. Calli induction depends on the 2-iP and 2.4 D ratio. The 20.0µM of 2.4-D and 20.0µM of 2-iP combination caused the highest embryogenic calli induction rate.

Visando a alcançar alta eficiência na indução de calos a partir de explantes foliares de plantas matrizes de C. arabica com alta heterozigose, por meio da embriogênese somática indireta, foram instalados três experimentos. O primeiro experimento foi conduzido em esquema fatorial 2x5, constituído de dois meios de cultura (BOXTEL & BERTHOULY, 1996 e TEIXEIRA et al., 2004) e cinco genótipos de C. arabica. No segundo experimento, foi avaliado o potencial de produção de calos embriogênicos em 10 genótipos, sendo cada genótipo considerado como um tratamento e, no terceiro experimento, foram avaliadas as variações nas concentrações de 2.4-D (2,5 e 20µM) e 2-iP (2,5 e 20µM) nos meios primário e secundário de TEIXEIRA et al. (2004). As culturas foram mantidas a 25oC, sob obscuridade. Para os genótipos 2.2 e 7.2, verificou-se a superioridade do meio de cultura Teixeira et al. (2004) em relação ao meio BOXTEL & BERTHOULY (1996). No genótipo 4.2, observou-se o comportamento inverso, ou seja, a superioridade do meio BOXTEL & BERTHOULY (1996). Os genótipos 3.0 e 5.0 apresentaram o mesmo comportamento em ambos os meios de cultura estudados, evidenciando que a produção de embriões somáticos é fortemente dependente do genótipo. A indução de calos depende da relação de 2-iP e 2.4-D. A combinação de 20.0µM of 2.4-D e 20.0µM of 2-iP promoveu a maior porcentagem de indução de calos embriogênicos.

Toxicidade de produtos fitossanitários utilizados em citros para Apis mellifera / Toxicity of pesticides used in citrus crop to Apis mellifera

O objetivo deste trabalho foi avaliar a toxicidade de inseticidas/acaricidas utilizados em cultura de citros para operárias africanizadas de Apis mellifera Linnaeus. A exposição das abelhas aos compostos foi realizada usando-se técnicas de pulverização, contaminação da dieta e contato em superfícies tratadas (folhas de citros e placas de Petri), empregando-se asdoses máximas recomendadas para a cultura. Os bioensaios foram realizados em laboratório a 25±2°C, UR 70±10% efotofase de 12h, sendo os dados de mortalidade submetidos à análise estatística, e as médias comparadas por contraste, obtendo-se grupos de efeitos semelhantes. Independente do modo de exposição, o acefato foi extremamente tóxico, matando mais de 90,0% das abelhas 24h após a aplicação. Os produtos espirodiclofeno e piriproxifem, quando aplicados diretamente sobre as abelhas, causaram níveis de mortalidade de 11,0 e 15,0%, respectivamente; os compostos buprofezina, enxofre e tetradifona apresentaram níveis de mortalidade ainda menores, com média de 5,0% entre eles. Para os ensaios decontaminação de superfície (folhas de citros e placas de Petri) e contaminação de alimento, foram obtidos dois grupos de toxicidade, um grupo somente com acefato e outro, com buprofezina, enxofre, espirodiclofeno, piriproxifem, tetradifona e água. A mortalidade média para esse segundo grupo, após 96h do início da exposição, foi de 31,0; 8,3 e 15,7%, respectivamente, para cada método de contaminação.

The aim of this research was to evaluate the toxicity of several acaricides/insecticides used in Brazilian citrus crop to africanized workers of Apis mellifera Linnaeus. The expositionof honey bees to the chemicals was performed by direct spraying, contamination of food, and contact in treated surface (citrus leaves and Petri dishes), using recommended rates ofapplication. The assays were carried out at 25±2°C, RH 70±10%, 12h of photophase and the data was statisticallyanalyzed, with mean values of mortality being compared through cluster analysis. In all assays acephate was highly toxic, with mortality at 24 hours around 90.0%. When spirodiclofen and pyriproxyfen, was sprayed directly into the honeybees, they caused mortality levels of 11.0 and 15.0%, respectively; buprofezin, sulphur and tetradifon were less toxic, with mean mortality of 5.0% among these compounds. For the assays from contamination surface (citrus leaves and Petri dishes) and food, two groups of chemicals with the same toxiceffects were observed, one with acephate and other with buprofezin, sulphur, spirodiclofen, pyriproxyfen, tetradifon and water. The average mortality after 96 hours of exposition was 31.0; 8.3 and 15.7%, respectively, for each method of contamination.

Bioatividade do Bacillus thuringiensis var. kurstaki (Berliner, 1915) para adultos de Apis mellifera Linnaeus, 1758 (Hymenoptera: Apidae) / Bioactivity of Bacillus thuringiensis var. kurstaki (Berliner, 1915) to adults of Apis mellifera Linnaeus, 1758 (Hymenoptera: Apidae)

Avaliou-se a influência do Bacillus thuringiensis var. kurstaki (Berliner) sobre adultos de Apis mellifera Linnaeus. Os experimentos foram realizados em laboratório a 28 ± 2 °C, UR 70 ± 10 por cento e fotofase de 12 horas. B. thuringiensis foi aplicado com pulverização sobre adultos, e fornecido através de solução aquosa de mel a 50 por cento e em adição à pasta Cândi, utilizando o produto comercial Dipel® PM. Esse produto quando aplicado com pulverização ou incorporado à pasta Cândi ou à solução aquosa de mel provocou mortalidade de adultos de A. mellifera em todas as concentrações utilizadas, com exceção de 0,25 g de Dipel®/100 mL adicionado à solução aquosa de mel a 50 por cento. Ao ser incorporado à pasta Cândi, a CL50 correspondeu a 0,325 g e a CL90 2,127 g do B. thuringiensis var. kurstaki/60 g de pasta. Adicionado à solução aquosa de mel a 50 por cento, a CL50 foi de 1,403 g e a CL90 foi de 7,759 g do B. thuringiensis var. kurstaki/100 mL de solução. Sintomas de infecção pelo B. thuringiensis foram identificados nas abelhas adultas e através do isolamento obteve-se uma cultura dessa bactéria o que comprovou a patogenicidade para adultos de A. mellifera.

The effects of Bacillus thuringiensis var. kurstaki (Berliner) on adults of Apis mellifera Linnaeus were evaluated. The bioassays were carried out under controlled conditions at 25±2°C, RH 70±10 percent and 12-h photophase. Adults of A. mellifera were exposed to the commercial product Dipel® PM. The following methods were used: direct spraying; supplying Bt with honey aqueous solution; and by a Candy paste added to Bt. B. thuringiensis caused mortality on A. mellifera adults, independent of the method used, except at 0.25 g of Bt/100 mL added to the honey aqueous solution. B. thuringiensis added to the Candy paste showed CL50 and CL90 of 0.325 g and 2.127 g of the product to 60 g of Candy paste, respectively. Dipel® PM added to the honey aqueous solution showed CL50 and CL90 of 1.403 g and 7.759 g of B. thuringiensis/100 mL, respectively. Infection symptoms by B. thuringiensis were identified on bees adult and by isolation of this bacterium, the toxicity of B. thuringiensis on A. mellifera was confirmed.